Acetylcholinesterase (AChE) can be released in the perfusate of rat caudate nucleus (CN) slices by two different modes of stimulation, with electrical stimulation at 5 Hz and with high concentrations of K+ (105 mM) using K+-propionate. We were unable to demonstrate AChE release with lower K+ concentrations (50 mM); however, at this concentration the drop in AChE activity seen in resting conditions was prevented. Practically all (95%) cholinesterase (ChE) found in the rat CN is AChE, which is represented by two major molecular forms (4S and 10S). In the perfusate, only AChE activity could be detected. A comparison of acetylcholine (ACh) and AChE release showed that maximal 3H-outflow and AChE release occurred at the same frequency (5 Hz), but the onset of AChE release was delayed. With high K+ (105 mM) depolarization, AChE release started after termination of the stimulation and continued for at least 50 min. These findings are consistent with the view that soluble form(s) of AChE can be slowly released from neurons under specific conditions of depolarization. In the caudate of the rat, the most likely sites for this release are processes of cholinergic interneurons. A hypothesis of AChE release is presented, and possible physiological and pathological implications of such a mechanism are discussed.